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Detection kit and application thereof

A detection reagent and kit technology, applied in the field of biological detection, can solve the problems of high price, cumbersome operation process, long time consumption, etc., and achieve the effect of rapid sensitivity

Active Publication Date: 2016-04-27
LUOYANG PULIKE WANTAI BIOTECH +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the methods for detecting these diseases include virus isolation and identification and serum examination, but the operation process is cumbersome and time-consuming; while PCR / RT-PCR detection is sensitive, but requires specialized instruments and professional operators. Not suitable for grassroots or on-site rapid detection
[0007] Although the prior art also has a single colloidal gold detection test strip for detecting the respective antigens, the price is expensive or the detection sensitivity is low

Method used

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  • Detection kit and application thereof
  • Detection kit and application thereof
  • Detection kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0034] As an embodiment of the present invention, the method includes: inserting the collected microbial swab into the sample processing tube, dissolving the sample in the solution as much as possible, adding the processed sample dropwise to the detection test strip In the center of the hole, after 10 minutes or 30 minutes, the results are judged: (1) If there is no band at the quality control line, no matter whether there is a band at the detection line, the detection process is invalid; (2) There is a band at the quality control line If there is a band at the detection line, it is positive, otherwise it is negative, that is, the presence or absence of the quality control line determines whether the detection process is effective. If there is no band at the quality control line, no matter whether there is a band at the detection line The detection process is invalid. If there is a band at the quality control line, it is positive if there is a band at the detection line, otherw...

Embodiment 1

[0041] Example 1 Preparation, purification, identification and testing of anti-porcine epidemic diarrhea virus monoclonal antibodies PEDV-McAB1 and PEDV-McAB2

[0042] 1.1 Preparation and purification of monoclonal antibody against porcine epidemic diarrhea virus

[0043] The porcine influenza virus HN1301 strain was purified according to the method of Zhang Liyan et al. (Zhang Liyan. Development of porcine epidemic diarrhea virus monoclonal antibody. Sichuan Agricultural University. Master's thesis), immunized mice, and obtained 2 hybridoma cell strains. Two monoclonal antibodies were prepared respectively.

[0044]The two monoclonal antibodies were prepared according to Chen Dan et al. ) operating method for purifying monoclonal antibodies by octanoic acid-ammonium sulfate combined precipitation method to purify anti-porcine epidemic diarrhea virus monoclonal antibodies PEDV-McAB1 and PEDV-McAB2 respectively.

[0045] 1.2 Identification of monoclonal antibodies against por...

Embodiment 2

[0087] Preparation and application of embodiment 2 kit

[0088] 2.1 Colloidal gold detection test strip of the kit

[0089] 2.1.1 Preparation and application of the colloidal gold detection test strip of the kit

[0090] The kit includes colloidal gold detection test strips, sample processing solution (i.e. pH7.4 phosphate buffer containing 2% CHAPS), sample processing tube, sample preservation solution, sample preservation solution (i.e. pH7.4 phosphate buffer solution) in the sample preservation bottle, wherein the preparation of the colloidal gold detection test strip is as follows:

[0091]Prepare colloidal gold solution according to the colloidal gold preparation method that China CN101614737A establishes, and mark respectively monoclonal antibody PEDV-McAB2 (the anti-porcine epidemic diarrhea virus monoclonal antibody PEDV-McAB2 that embodiment 1 prepares, use pH7.4 phosphate buffer saline diluted to a labeled concentration of 250 μg / ml), monoclonal antibody TGEV-McAB4...

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Abstract

The invention provides a detection kit containing two anti porcine epidemic diarrhea virus monoclonal antibodies PEDV-McAB1 and PEDV-McAB2, two anti porcine transmissible gastroenteritis virus monoclonal antibodies and two anti porcine rotavirus monoclonal antibodies; the kit can be applied for simultaneous detection of porcine epidemic diarrhea viruses, porcine transmissible gastroenteritis viruses and / or porcine rotaviruses with non-diagnostic purpose, moreover, the detection sensitivity of the kit for simultaneous detection of two or three kinds of viruses is higher than that of single detection of one kind of virus, and false positive results are avoided.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a detection kit and its application. Background technique [0002] Porcine epidemic diarrhea (porcineepidemicdiarrhea, PED) is an acute, highly contagious infectious disease caused by porcine epidemic diarrhea virus (porcineepidemicdiarrheavirus, PEDV) of the family Coronaviridae. It is characterized by diarrhea and dehydration, and pigs of all ages are susceptible. Among them, the incidence rate of suckling piglets, shelf pigs or fattening pigs after infection can reach 100%, and suckling piglets suffer the most. In recent years, since the winter of 2010, porcine epidemic diarrhea has broken out in most parts of my country, and then it has also broken out in South Korea, Japan, Taiwan and other regions on a large scale. In the first half of 2013, the disease also broke out in North America and spread rapidly, causing serious losses to the global breeding industry...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/569G01N33/558
CPCG01N33/558G01N33/56983G01N33/577G01N2333/14G01N2333/165G01N2333/17
Inventor 田克恭王莹
Owner LUOYANG PULIKE WANTAI BIOTECH
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